Team:Cambridge/Bioluminescence/G28

From 2010.igem.org

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{{:Team:Cambridge/Templates/rightpic|src=Jungle_book.jpg|300px|right|G28 illuminating the Jungle Book}}
{{:Team:Cambridge/Templates/rightpic|src=Jungle_book.jpg|300px|right|G28 illuminating the Jungle Book}}
{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}
{{:Team:Cambridge/Templates/rightpic|src=Space_invader.jpg|400px|right|G28 on a 96 well plate}}
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James Slock from King's College, PA kindly provided us with plasmids carrying the genes responsible for bioluminescence in ''V. fischeri''. Using Long-Range PCR, we extracted these genes and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], The lux operon in ''V. fischeri'' is under tight quorum sensing control. In the absence of LuxR and AHL the Lux genes are virtually inactive. In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to produce an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct G28. It caused bright and reproducible light output in the transformed E.coli Top10 cells. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using this strain.  
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James Slock from King's College, PA kindly provided us with plasmids carrying the genes responsible for bioluminescence in ''V. fischeri''. Using Long-Range PCR, we extracted these genes and assembled them into a new operon. As described in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background Background section], The lux operon in ''V. fischeri'' is under tight quorum sensing control. In the absence of LuxR protein and AHL the Lux genes are virtually inactive. In order to relieve this control, we used [https://2010.igem.org/Team:Cambridge/Gibson/Introduction Gibson Assembly] to produce an operon consisting of Lux C, D, A, B, E (in this order, reflecting ''V. fischeri'') under the arabinose-induced promoter pBAD ([http://partsregistry.org/Part:BBa_I0500 BBa_i0500]). We called this construct G28. It caused bright and reproducible light output in the transformed E.coli Top10 cells. This new BioBrick ([http://partsregistry.org/Part:BBa_K325909 BBa_K325909]) can be used as an arabinose->light device and is a very useful part if the aim is to get a high bacterial light output. Many of the images in our [https://2010.igem.org/Team:Cambridge/Photos Photo Gallery] were created using Top10 cell transformed with this part.  
=h-ns mutants=
=h-ns mutants=

Revision as of 22:16, 25 October 2010